Dear Tim:
Depending on what are you going to do after recovery of the
protein from the PVDF, the different procedures may work or may not. I
tried different solvents ( acetonitrile, isopropanol, TFA, Buffers,
detergents ,etc and mixtures of them ) and no succesful at all ( at least in
my hands ) when I checked the solution by AAA. But, what I routinely do
with secuenced PVDF bands is aminoacid analysis, to know how much protein
was/is there. For that purpose, I put the band directly on the 420 ABI glass
slide, covering it with a round plastic net ( I have the number spare part
). The machine offers automatic hydrolisis but in case there is not this
possibility, the PICO-TAG station works fine and after gas phase
hydrolisis, you extract the aminoacids with 100 ul of ClH 6 N first and
100 ul of a ACN-water-TFA 0,1% ( 100:50:20 ) in a second step. , vortexing
thoroughly .
Hope that it helps
Susana
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Susana Linskens
LANAIS-PRO
Junin 956
1113 Buenos Aires - ARGENTINA
linskens@qb.ffyb.uba.ar
FAX: 54-11-4508-3652
Phone: 54-11-4508-3651
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